Construction and characterization of an NaCl-sensitive mutant of Halomonas elongata impaired in ectoine biosynthesis.
Construction and characterization of an NaCl-sensitive mutant of Halomonas elongata impaired in ectoine biosynthesis.
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DOI：
10.1111/j.1574-6968.1998.tb12960.x
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发表时间：
1998-04
影响因子：
2.1
通讯作者：
K. Göller;Alexandra Ofer;E. Galinski
中科院分区：
文献类型：
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作者：
K. Göller;Alexandra Ofer;E. Galinski
使用转座子诱变，我们产生了盐敏感的突变体的嗜盐真杆菌长盐单胞菌受损的生物合成兼容的溶质四氢嘧啶。细胞质溶质含量的HPLC测定显示四氢嘧啶的生物合成前体L-2，4-二氨基丁酸的积累。未检出埃克托因和羟基埃克托因。该突变体不能在NaCl浓度超过4%的基本培养基中生长。然而，当补充有机渗透剂，在高盐度介质（15%和更高）中生长的能力得到恢复。我们克隆并测序了转座子插入H.长体染色体与已知蛋白质的序列比较显示，突变基因的L-2，4-二氨基丁酸乙酰转移酶的四氢嘧啶的生物合成途径在嗜盐海洋球菌的显着相似性。对PCR产物的分析表明，四氢嘧啶生物合成基因（ectABC）遵循与M相同的顺序。嗜盐菌
Using transposon mutagenesis we generated a salt-sensitive mutant of the halophilic eubacterium Halomonas elongata impaired in the biosynthesis of the compatible solute ectoine. HPLC determinations of the cytoplasmic solute content showed the accumulation of a biosynthetic precursor of ectoine, L-2,4-diaminobutyric acid. Ectoine and hydroxyectoine were not detectable. This mutant failed to grow in minimal medium with NaCl concentrations exceeding 4%. However, when supplemented with organic osmolytes, the ability to grow in high-salinity medium (15% and higher) was regained. We cloned and sequenced the regions flanking the transposon insertion in the H. elongata chromosome. Sequence comparisons with known proteins revealed significant similarity of the mutated gene to the L-2,4-diaminobutyric acid acetyltransferase from the ectoine biosynthetic pathway in Marinococcus halophilus. Analysis of a PCR product demonstrated that the ectoine biosynthetic genes (ectABC) follow the same order as in M. halophilus.